CN108403690A - It is a kind of inhibit melanoma cells proliferation drug and its application - Google Patents
It is a kind of inhibit melanoma cells proliferation drug and its application Download PDFInfo
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- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
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- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
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Abstract
The invention belongs to biomedicine fields, are related to a kind of drug of inhibition melanoma cells proliferation, active constituent includes 2,3 istains and/or 5 fluorouracils.2,3 istains promote B16 Apoptosis, inhibit melanoma cell growth by lowering 2 protein expressions of bcl, and do not reduce immunity, it has no toxic side effect, 2,3 istains can be by B16 cell-cycle arrests in the S phases, that is DNA synthesizes the phase, and there are sub G1 apoptotic peaks, apoptotic peak obviously increases after being combined with 5 fluorouracils, and Cycle Arrest effect becomes apparent from；To various immunocyte numbers without obvious inhibiting effect in 2,3 istain bodies, and certain 5 fluorouracil immunosuppressive effect of mitigation is shown after being shared with 5 fluorouracils.This provides a kind of new solution for melanoma cells more effectively treatment.
Description
Technical field
The invention belongs to biomedicine field, especially melanoma therapy field, specially a kind of inhibition melanoma
The drug of cell Proliferation and its application.
Background technology
Malignant mela noma (Malignant melanoma, MM) is referred to as disliked black, is most common evil in worldwide
Property one of tumour, it originates from human nerve's ridge melanocyte, and by melanocyte vicious transformation froing, it, which has, highly invades
It attacks, the characteristic of height transfer.According to incompletely statistics, the incidence of human malignant's melanoma accounts for mankind's entirety malignant tumour
3%, and the incidence of human skin malignant mela noma accounts for the 7%-20% of mankind's whole malignant tumour of skin, and it is sent out
Sick rate is still increased with 4%-6% speed every year.
Up to the present, include mainly operative treatment, radiotherapy, chemistry to the therapy of human malignant's melanoma
Treatment, biological therapy and tumor vaccine therapy.And drug therapy or topical medications can carry for part melanoma patients
For a kind of preferably prevention and treatment effect, reach the therapeutic purposes for extending life cycle, improving the quality of living.Therefore, it finds high
The drug candidate for imitating the melanoma of low toxicity is particularly important.The drug having disclosed have PAK1 (CN201380014567.8),
Lipopeptide derivatives (CN201410370095.6), chlorogenic acid (CN201510079033.4), all-trans retinoic acid (ATRA) and acid
Mould element -8-0- β-D- glucopyranosides use in conjunction (CN201410475269.5) etc..But existing medication effect is not
Stablize, and normal tissue and immune system also have lethal effect, often will appear leucocyte caused by being reduced by immunosupress,
The toxic side effects such as blood platelet reduction and loss of appetite, the nausea and vomiting of digestive system, these side effects and drug resistance of tumor
The factors such as generation can generate larger limitation to the application of chemotherapy.Therefore, on the basis of not reducing immunity, seek
The new anti-melanin tumor medicine having no toxic side effect is imperative.
2,3-indolinedione (isatin, ISA), molecular weight 147.13 are a kind of naturally occurring Benzazole compounds, are
It is widely present in the endogenous active substance of animal and human body.2,3-indolinedione chemical constitution is clear, is that lobster sustains life
Necessary material.Find that 2,3-indolinedione is present in urine extract in human body for the first time, with hindbrain and cardiac muscle in have also discovered this
Substance especially has higher distribution at positions such as intracerebral hippocampus.With the further investigation to its pharmacological action, it is found that it is extensive it has
Biological activity, there are some researches prove 2,3-indolinedione have the effects that explicitly ease pain, anti-inflammatory, antiviral, antidepression, and
It is apparent to kinds of tumors modelling effect.The connected dimeric indole structure of carbon bridge is the key that antitumor activity.2,3- istains are
The precursor of anti-cancer drugs indigo red chemical dimer structure, and structure is simpler compared with indigo red, overcomes steric hindrance, and exist
1 and 2 chemical allosteric group, increases the penetration capacity to mucous epithelium, thus is more easy to be absorbed.In terms of structure-activity relationship,
2,3- istains have the condition of anticancer and anti-cancer.Wherein, Chinese invention patent CN200410023871.1 discloses 2,3-
Istain can inhibit PCNA, Bcl-2 protein expression in S180 tumor tissues, can promote Human neuroblastoma cell apoptosis and inhibit it
Proliferation；2,3-indolinedione has antitumor action, mechanism related with tumor cell proliferation, inducing apoptosis of tumour cell is inhibited.
Studies in China personnel in 2007 demonstrate in 2,3- istains body has work to SH-SY5Y, H22, EAC transplanted tumor model
With.2014 researchers have shown that 2,3-indolinedione can lower the expression of human breast cancer cell (MCF-7) Bcl-2 albumen, should
Mechanism may be related with 2,3- istain inducing apoptosis of tumour cell.Meanwhile the result of study finds that 2,3-indolinedione can also be led
Cause cell-cycle arrest, mechanism unknown.Also literature has shown that, ISA can inhibit the proliferation of SH-SY5Y neuroblastomas simultaneously
It is apoptosis-induced.2016, studies in China personnel had found that ISA can inhibit the invasion of SH-SY5Y again.
However, not inhibiting the document report of melanoma cells proliferation about 2,3-indolinedione still up to now, do not have more
There is the document report of anti-melanin oncocyte after being combined it with other anticancer drugs.
Invention content
The purpose of the present invention is to provide a kind of drugs of inhibition melanoma cells proliferation and its application, the drug to pass through
Melanoma Bcl-2 protein expressions are lowered to promote B16 Apoptosis, inhibit melanoma cell growth, and are not reduced immune
Power has no toxic side effect, and has new drug development value in treatment malignant mela noma.Therefore, the purpose of the present invention is to be achieved
's：A kind of drug inhibiting melanoma cells proliferation, the drug include active constituent and pharmaceutically available carrier and/or tax
Shape agent, wherein active constituent include 2,3-indolinedione.
The present invention is also combined by body outer cell line and Inoculation scale-model investigation 2,3- istains and 5 FU 5 fluorouracil
To inhibiting the effect of mouse melanin tumor cell proliferation, body outer cell line is as a result, it has been found that 2,3-indolinedione can be by cell-cycle arrest
In the S phases, and there are sub-G1 apoptotic peaks, positive control medicine 5 FU 5 fluorouracil also can arresting cell cycle, but do not have apoptosis
Peak, apoptotic peak obviously increases after combination, and Cycle Arrest effect becomes apparent from, and inhibits the effect enhancing of melanoma cells proliferation, this
It obtains more effectively treating for melanoma cells and provides a kind of new solution.Inoculation model finds 2,3- indoles
Quinone can obviously inhibit the growth of melanoma, to various immunocyte numbers without obvious inhibiting effect, and with 5 FU 5 fluorouracil
Certain mitigation 5 FU 5 fluorouracil immunosuppressive effect is shown after combination.Therefore, the purpose of the present invention can also be real in this way
It is existing：Active constituent in the drug for inhibiting melanoma cells proliferation includes 2,3- istains and 5 FU 5 fluorouracil.At this
It invents in preferred technical solution, the active constituent in the pharmaceutical composition is made of 2,3-indolinedione and 5 FU 5 fluorouracil.
Carrier and/or excipient of the present invention include but not limited to：Brine, glucose, water, dimethyl sulfoxide (DMSO) and
A combination thereof；Preferably dimethyl sulfoxide (DMSO).
The present invention shows that 2,3-indolinedione can induce B16 Apoptosis by each group Apoptosis testing result, and is in agent
Measure dependence.Preferably, in the pharmaceutical composition of the present invention for inhibiting melanoma cells proliferation, matched with dimethyl sulfoxide (DMSO)
The 2,3-indolinedione mother liquor for setting 100 μM dilutes mother liquor into a concentration of 100-800 μ with 1640 culture mediums containing 10% fetal calf serum
M；Preferably 400 μM；The 5 FU 5 fluorouracil mother liquor that 100mM is configured with dimethyl sulfoxide (DMSO), with 1640 trainings containing 10% fetal calf serum
It supports base and dilutes mother liquor into a concentration of 50-350 μM；Preferably 200 μM.
The invention also provides 2,3- istains promote B16 Apoptosis preparing downward Bcl-2 protein expressions and will
Application of the cell-cycle arrest in drug of the S phases to inhibit melanoma cell growth.The invention also provides said medicines
Application in preparing the drug for inhibiting melanoma cells proliferation also has and mitigates immunosuppressive effect, especially mitigates 5-
Fluorouracil immunosuppressive effect.
The invention also discloses a kind of inhibition melanoma cells proliferation and mitigate immunosuppressive method, said medicine
Intervene in vitro and/or in vivo melanoma cells.The melanoma cells are mouse melanin tumor cells.
Compared with prior art, the drug of the present invention for inhibiting melanoma cells proliferation has the following advantages and shows
Write progress：2,3-indolinedione promotes B16 Apoptosis, inhibits melanoma cell growth by lowering Bcl-2 protein expressions,
And immunity is not reduced, it has no toxic side effect, 2,3-indolinedione can be by B16 cell-cycle arrests in the S phases, i.e. DNA synthesizes the phase, and goes out
Existing sub-G1 apoptotic peaks, apoptotic peak obviously increases after being combined with 5 FU 5 fluorouracil, and Cycle Arrest effect becomes apparent from；2,3- istains
In vivo to various immunocyte numbers without obvious inhibiting effect, and show that mitigation 5- fluorine urine is phonetic after being combined with 5 FU 5 fluorouracil
Pyridine immunosuppressive effect.
Description of the drawings
Fig. 1 is cell number testing result, and (A) is FCM analysis image, and (B) is the statistical chart of cell number.
Fig. 2 is the cell cycle as a result, (A) is flow cytomery image, and (B) is each period proportional statistical result.
Fig. 3 is Apoptosis as a result, (A) is flow cytometer image, and (B) is Q3 quadrant statistical results.
Fig. 4 is Bcl-2 average fluorescent strengths, and (A) is streamed image, and (B) is statistical chart.
Fig. 5 is tumor growth curve.
Fig. 6 is knurl weight result.
Fig. 7 is each group's immunocyte number in spleen, and (A) is total leukocyte number statistical figure, and (B) is neutrophil leucocyte system
Meter figure, (C) is bone-marrow-derived lymphocyte number statistical figure, and (D) is NK cell number statistical charts.
Specific implementation mode
The specific implementation mode of the present invention is described further below in conjunction with the accompanying drawings.
1, body outer cell line is tested：Murine melanoma (B16) cell is provided simultaneously by Taishan Hospital's immunological investigation
It preserves.B16 cell recoveries are placed in 1640 culture mediums containing 10% fetal calf serum and are cultivated, are received when growing into exponential phase
Collect cell, and is uniformly taped against on 24 orifice plates, plating density 1*105A/hole carries out Apoptosis, cell cycle and detection
Bcl-2 is tested, and studies the relationship and relevant molecule mechanism of 2,3-indolinedione and B16 cell cycles.
Embodiment 1-4
Embodiment 1-4 is illustrating body outer cell line experimental data：
B16 cell lines are divided into four groups：Control group, 2,3- istains group, 5 FU 5 fluorouracil group and drug combination group.Every group
Six holes handle 48h.With flow cytometer, the holes constant volume 300ul/, identical speed, the fixed holes receipts sample time 120s/ carry out cell
Number detection, cell cycle detection, apoptosis detection, Bcl-2 detection of expression.
Embodiment 1 is control group, and 1%DMSO is added.
Embodiment 2 is 2,3-indolinedione group, and 2,3-indolinedione is configured to the mother of 100mM with dimethyl sulfoxide (DMSO) (DMSO)
Liquid：The relative molecular mass of 2,3-indolinedione is 147, so weighing 14.7mg2,3- istains are dissolved, gained with 1mlDMSO
Solution is the 2,3- istain mother liquors of 100mM.With 1640 culture mediums containing 10% fetal calf serum be diluted to respectively 200uM,
400uM, 800uM are spare.
Embodiment 3 is 5 FU 5 fluorouracil group, and 5 FU 5 fluorouracil is dissolved into the mother liquor of 100mM with DMSO：Weigh 13mg5-
Fluorouracil powder, is dissolved with 1mlDMSO, and acquired solution is the 5 FU 5 fluorouracil mother liquor of 100mM.With containing 10% fetal calf serum
1640 culture mediums to be diluted to 200uM spare.
Embodiment 4 is drug combination group, and 400uM 2,3-indolinediones are added and 200uM 5 FU 5 fluorouracils (contain 1%
DMSO)。
Test case 1-4
Test case 1-4 is respectively cell number detection, cell cycle detection, Apoptosis detection and Bcl-2 mean fluorecences
Intensity detection, test result are shown in attached drawing 1-4.Statistical chart is compared examined using t two-by-two using the mapping of Prism7 softwares and statistical analysis
It tests, " * " represents P 0.05, and " * * " represents P 0.01, and " * * * " represents P 0.001, and " * * * * " represents P 0.0001.
Test case 1：Cell number detects
After pharmaceutical intervention 24 hours, culture medium is discarded, twice of PBS rinses are added, 0.25% trypsase is added and (contains
EDTA), 37 DEG C of digestion 2min pay attention to observing cellular morphology, in the experimental result of the fixed cell number received and collected in the sample time
See Fig. 1.The results show that the cell number of embodiment 2-4 (2,3-indolinedione group, 5 FU 5 fluorouracil group and two medicine combination groups) is apparent
Less than embodiment 1 (control DMSO groups), and drug effect does not reduce after combination, shows 2,3-indolinedione, 5 FU 5 fluorouracil and the two connection
It is proliferated with there is significantly inhibiting mouse melanin tumor cell.
Test case 2：Cell cycle is detected
It after drug-treated, is dyed by PI, PI is combined with intracellular DNA, is carried out according to the content distribution situation of DNA thin
The analysis in born of the same parents' period and apoptosis.After PI is dyed, it is assumed that the fluorescence intensity of G0/G1 phases is 1, and the cell of apoptosis occurs due to thin
Karyon can be shunk and the fragmentation of DNA causes some DNA fragmentations that can be lost during dyeing, thus it is shown that glimmering
Luminous intensity is less than 1 peak, and the peaks sub-G1, that is, Apoptosis peak will be shown as on the fluorogram of flow cytometer detection.As a result
See Fig. 2, (2,3-indolinedione group) cell cycle S of embodiment 2 phase ratio is significantly raised, and sub-G1 apoptotic peaks occurs, embodiment 3
The S phases of (positive control medicine 5 FU 5 fluorouracil) also obviously increase, but do not have apoptotic peak, and embodiment 4 (drug combination group) is withered
Peak is died to obviously increase.The result shows that：2,3-indolinedione can be by B16 cell-cycle arrests in the S phases, i.e. DNA synthesizes the phase, and occurs
Sub-G1 apoptotic peaks, another S phases block antitumor drug-positive control medicine 5 FU 5 fluorouracil also can arresting cell cycle,
But there is no apoptotic peak, the period, blockage effect became apparent from after combination, enhanced the inhibiting effect being proliferated to melanoma cells.
Test case 3：Apoptosis detects
After drug-treated 24 hours, each group cell is collected in respectively in 96 orifice plates, is numbered according to grouping, used
Annexin v, 7-aad apoptosis detection kits must use stream according to illustrating to be dyed in 1 hour after dyeing
Formula cell instrument is detected, and the quadrant of main detection Annexin v+ and 7-aad-, this quadrant are represented into Apoptosis process
Cell.Experimental result is shown in Fig. 3, the Annexin v+ of the B16 cells of the 2,3-indolinedione processing of 200uM, 400uM, 800uM concentration
And the ratio of 7-aad- quadrants is apparently higher than control group, this illustrates that the 2,3-indolinedione of various concentration can promote B16 cells
Apoptosis, and be in dose dependent.
Test case 4：Bcl-2 average fluorescent strengths detect
After carrying out pharmaceutical intervention 24 hours by grouping, cell is collected in 96 orifice plates, by flow cytometry antibody labeled cells
The method of molecule is marked in core, after use flow cytomery.The expression height of institute's mark molecule can be glimmering with this
The average fluorescent strength of photoactivated antibody indicates that average fluorescent strength is higher, then the single intracellular Bcl-2 expression of explanation is higher, on the contrary
It is then relatively low.Experimental result is shown in Fig. 4, the average fluorescent strength of embodiment 2 (2,3-indolinedione group) Bcl-2 is substantially reduced, and is used
Prism7 softwares are mapped and are counted, and average fluorescent strength has pole significant difference P 0.001 between two groups.
Vitro Experimental Results show that 2,3- istains have apparent inhibition growth to mouse melanin tumor cell, promote apoptosis
Effect.2,3-indolinedione has obvious inhibiting effect to B16 cell Proliferations in vitro, it inhibits certain tumours thin with document report
The proliferation of born of the same parents such as SH-SY5Y, the female tumor of mouse NE-115 nerves, S180, H22 etc. match.Flow cytomery finds 2,3- Yin
Diindyl quinone can promote the apoptosis of B16 cells, cell cycle result to also show the ratio apparent increase of S phases by lowering the expression of Bcl-2,
To promote Apoptosis while arresting cell cycle, inhibit the proliferation of B16 cells.Bcl-2 is the anti-apoptotic proteins of broad sense,
It can be by inhibiting Apoptosis to participate in the generation of tumour.The above results illustrate that 2,3- istains have melanoma in vitro
Effect, this effect can inhibit B16 cell Proliferations with 2,3-indolinedione, arresting cell cycle, inducing cell apoptosis, lower Bcl-2
Expression it is related.
2, internal cell line assay：This experiment is purchased from Beijing China using the C57BL/6 inbred mouses of female 6-8 week old
Fukang bio tech ltd.B16 cells are seeded in C57BL/6 mouse backs by the numbers of 5*105/ only, are randomly divided into
Four groups, tumour growth situation is recorded daily, and gavage is put to death after two weeks, tumor tissues of weighing, each group's immunocyte such as detection spleen
Number of variations.
Embodiment 5-8
Embodiment 5-8 is illustrating experiment in vivo data.
Embodiment 5 is control group, prepares the tragacanth solution of 1.25g/mL.
Embodiment 6 is 2,3-indolinedione group, prepares the 2,3-indolinedione suspension of 100mg/kg：The C57 mouse of 6-8 week old
Only, so the quality of 2,3-indolinedione needed for every mouse is 2mg, every gavage volume is 200ul to general 20g/, therefore matches 2,
A concentration of 2mg/200ul of 3- istain suspensions, i.e. 2g/200ml.2g2 accurately is weighed, 3- istain powder is all added to
In the bottle of 1.25g/mL tragacanths for filling 200ml, mix well, it when necessary can ultrasound.
Embodiment 7 is 5 FU 5 fluorouracil group, prepares the 5 FU 5 fluorouracil solution of 25mg/kg, and Mice Body weighs about 20g/ only, institute
It is 0.5mg, every gavage 200ul with every required 5 FU 5 fluorouracil, then the concentration of solution is exactly 0.5mg/200ul, i.e.,
500mg/200ml.5 FU 5 fluorouracil powder 500mg accurately is weighed, is transferred completely into the bottle for filling 200ml tragacanth solution
In, 37 DEG C of water bath sonicators are put into, waits for that graininess 5 FU 5 fluorouracil is completely dissolved, mixes well spare.
Embodiment 8 is drug combination group.
Test case 5-7：
Test case 5-7 is respectively that tumour growth, knurl weight and each internal organs immunocyte number difference, test result are shown in attached drawing 5-
7.Statistical chart is compared examined using t two-by-two using the mapping of Prism7 softwares and statistical analysis, and " * " represents P 0.05, and " * * " is represented
P 0.01, " * * * " represent P 0.001, and " * * * * " represents P 0.0001.Circle on line chart in Fig. 5-7 represents western yellow alpine yarrow
Glue control group, square represent 2,3-indolinedione group, and triangle represents 5 FU 5 fluorouracil group, and up-side down triangle represents 2,3-indolinedione
With 5 FU 5 fluorouracil combination group
Test case 5：Tumor growth curve is tested
With electronic vernier caliper, automatic reading.Major diameter and minor axis are recorded when measurement, and tumour body is subsequently calculated according to formula
Product.Tumor growth curve result is shown in that Fig. 5, ordinate are tumor size, with " major diameter x minor axis2/ 2 " calculate.It is empty to record result display
One group of mouse of 1% tragacanth of white control group, that is, gavage occurs tumour earliest and the speed of growth leads over always other three groups.
Test case 6：Knurl weight result
Mouse is put to death in the 15th day of Mice Inoculated melanoma, completely strips out careful weighing after tumour, acquired results
See Fig. 6, (A) is the tumour of each group mouse, and every group is a line, is embodiment 5-8 successively from top to bottom.For convenience of comparison, put
When every group put by descending order.(B) it is knurl weight statistical chart, the results show that embodiment 5 is blank control group tumour weight
Amount is significantly greater than embodiment 6-8, and indifference between tri- groups of embodiment 6-8.This, which directly demonstrates 2,3- istains, to inhibit
The effect of mouse melanoma, and distich with drug 5 FU 5 fluorouracil without influence.
Test case 7：Each internal organs immunocyte number difference
Spleen is the maximum organ of immune organ and lymphatic system important in animal body, it can to whole body respectively by
Immunocyte is transported to the position of infection, if spleen is destroyed or influenced, this is a heavy losses to the immune system of body.
The cell colony of the CD45 positives is considered total leukocyte；Under the premise of the CD45 positives, the cell of the CD3 positives is total T lymphs
Cell；Under the premise of the CD3 positives, the cell colony of the CD4 positives and the CD8 positives respectively represents CD4+T lymphocytes and CD8+T
Lymphocyte；The B220 positives are bone-marrow-derived lymphocytes；The cell of the NK1.1 positives is considered NK cells；In the premise of the CD45 positives
Under, a group cell of CD11b and the bis- positives of Ly6G is neutrophil leucocyte.Experimental result is shown in Fig. 7, the results showed that, use 2,3-
The mouse of the inoculation B16 cells of istain intervention, its leucocyte total amount, neutrophil numbers, bone-marrow-derived lymphocyte number after execution
Compared with embodiment 5 without significant change, this illustrates that 2,3-indolinedione does not interfere with the normal immune response of body for mesh and NK cell numbers,
Immunocyte number in spleen does not change；And spleen total white blood cells, neutrophil numbers, B in 5 FU 5 fluorouracil group
Lymphocyte number and NK cell numbers decline compared with embodiment 5 and 6, and significant difference, this illustrates 5 FU 5 fluorouracil to machine
Body has apparent immunosuppressive action；The neutrophil numbers and NK cell number ratios embodiment 7 of 8 combination group of embodiment have bright
Aobvious to increase, compared with Example 5 without significant difference, this illustrates the substantially reduced 5 FU 5 fluorouracil of 2,3-indolinedione to spleen
The inhibiting effect of interior neutrophil leucocyte and NK cell numbers has also discovered combination group in the detection of leucocyte and bone-marrow-derived lymphocyte
Cell number risen than embodiment 7.It illustrates to show certain subtract after 2,3- istains are combined with 5 FU 5 fluorouracil
Light 5 FU 5 fluorouracil immunosuppressive effect.
2,3-indolinedione is as the endogenous active substance in organism, low self-evident, the experimental result of toxicity
It proves, 2,3-indolinedione plays the role of significantly inhibiting mouse melanoma in vivo, in this experiment, according to each in spleen
Number comparison of group's immunocyte after different pharmaceutical intervention, illustrates in 2,3-indolinedione body to various immunocyte numbers without bright
Aobvious inhibiting effect, and certain mitigation 5 FU 5 fluorouracil immunosuppressive effect is shown after being combined with 5 FU 5 fluorouracil.
As a kind of specific natural anti-cancer compound of structure, 2,3-indolinedione is in vivo and in vitro to murine melanoma
Cell growth has apparent inhibiting effect, and does not reduce immunity, has no toxic side effect, and after being combined with 5 FU 5 fluorouracil
Show certain mitigation 5 FU 5 fluorouracil immunosuppressive effect.So having new drug development valence in treatment malignant mela noma
Value.
Claims (10)
1. a kind of drug inhibiting melanoma cells proliferation, the drug available carrier comprising active constituent and pharmaceutically
And/or excipient, which is characterized in that the active constituent includes 2,3-indolinedione.
2. a kind of drug inhibiting melanoma cells proliferation according to claim 1, which is characterized in that in the drug
Active constituent include 2,3- istains and 5 FU 5 fluorouracil.
3. a kind of drug inhibiting melanoma cells proliferation according to claim 1, which is characterized in that in the drug
Active constituent be made of 2,3- istains and 5 FU 5 fluorouracil.
4. according to a kind of drug of any inhibition melanoma cells proliferation of claim 1-3, which is characterized in that described
Carrier and/or excipient include but not limited to：Brine, glucose, water, dimethyl sulfoxide (DMSO), and combinations thereof.
5. a kind of drug inhibiting melanoma cells proliferation according to claim 4, which is characterized in that the carrier
And/or excipient is dimethyl sulfoxide (DMSO).
6. a kind of drug inhibiting melanoma cells proliferation according to claim 5, which is characterized in that sub- with dimethyl
Sulfone configures 100 μM of 2,3-indolinedione mother liquor, and a concentration of 100-800 μ are diluted to 1640 culture mediums containing 10% fetal calf serum
M；The 5 FU 5 fluorouracil mother liquor that 100mM is configured with dimethyl sulfoxide (DMSO), 50- is diluted to 1640 culture mediums containing 10% fetal calf serum
350μM。
7. application of the drug described in claim 1-6 in preparing the drug for inhibiting melanoma cells proliferation.
8. application according to claim 7, which is characterized in that drug described in claim 1-6 can mitigate immunosupress.
9.2,3- istains promote B16 Apoptosis and by cell-cycle arrest in the S phases preparing downward Bcl-2 protein expressions
The application in drug to inhibit melanoma cell growth.
10. a kind of method inhibiting melanoma cells proliferation, drug described in claim 1-6 in external and/or dry in vivo
Pre- melanoma cells, the melanoma cells are mouse melanin tumor cells.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110694070A (en) * | 2019-11-27 | 2020-01-17 | 西安交通大学医学院第一附属医院 | Application of cytotoxic antitumor drug in preparation of tumor-related tissues |
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